Ontology highlight
ABSTRACT: Aims
Human embryonic stem cells (hESCs) can be used to generate scalable numbers of cardiomyocytes (CMs) for studying cardiac biology, disease modelling, drug screens, and potentially for regenerative therapies. A fluorescence-based reporter line will significantly enhance our capacities to visualize the derivation, survival, and function of hESC-derived CMs. Our goal was to develop a reporter cell line for real-time monitoring of live hESC-derived CMs.Methods and results
We used CRISPR/Cas9 to knock a mCherry reporter gene into the MYH6 locus of hESC lines, H1 and H9, enabling real-time monitoring of the generation of CMs. MYH6:mCherry+ cells express atrial or ventricular markers and display a range of cardiomyocyte action potential morphologies. At 20?days of differentiation, MYH6:mCherry+ cells show features characteristic of human CMs and can be used successfully to monitor drug-induced cardiotoxicity and oleic acid-induced cardiac arrhythmia.Conclusion
We created two MYH6:mCherry hESC reporter lines and documented the application of these lines for disease modelling relevant to cardiomyocyte biology.
SUBMITTER: Tsai SY
PROVIDER: S-EPMC7252441 | biostudies-literature | 2020 Mar
REPOSITORIES: biostudies-literature
Tsai Su-Yi SY Ghazizadeh Zaniar Z Wang Hou-Jun HJ Amin Sadaf S Ortega Francis A FA Badieyan Zohreh Sadat ZS Hsu Zi-Ting ZT Gordillo Miriam M Kumar Ritu R Christini David J DJ Evans Todd T Chen Shuibing S
Cardiovascular research 20200301 3
<h4>Aims</h4>Human embryonic stem cells (hESCs) can be used to generate scalable numbers of cardiomyocytes (CMs) for studying cardiac biology, disease modelling, drug screens, and potentially for regenerative therapies. A fluorescence-based reporter line will significantly enhance our capacities to visualize the derivation, survival, and function of hESC-derived CMs. Our goal was to develop a reporter cell line for real-time monitoring of live hESC-derived CMs.<h4>Methods and results</h4>We used ...[more]